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b-sitosterol, d-amyrin, d-amyrone, and quercetin phytochemicals in CQ. The
presence of ascorbic acid has been known to induce osteoblast specific gene
expression and proline and lysine hydroxylation in collagen.
98
Nanofibers
scaffolds of CQ prepared with hydroxyapatite and polycaprolactone have also
been reported to show beneficial effects in bone tissue regeneration.
99
The
3D printed b-tricalcium phosphate (TCP) loaded polydopamine loaded CQ
implants with improved in vivo new bone formation was developed by
Robertson and Bose.
100
Polydopamine showed increased proliferation and
alkaline phosphatase activity in osteoblast cell culture which was further
enhanced with the addition of CQ extract. The implants showed more new
bone growth compared to b-tricalcium phosphate. Figure 12.5 shows the
polydopamine and CQ coating to b-TCP scaffolds.
A novel herbal scaffold of 2% natural biopolymer alginate, O-carbox-
ymethyl chitosan, and herbal extract of CQ was developed by Soumya et al.
101
Initial cell attachment and proliferation of herbal extract was well spread
across the scaffold surf ace. The scaffold enhanced the activity of ALKP and
differentiation of human mesenchymal stem cells (hMSCs) to osteoblasts
with increased calcified mineral deposition. The stimulation of osteo-
blastogenesis is because of the presence of phytogenic steroids in CQ extract
and its action on estrogen receptors. The differentiation, proliferation of
osteoblast and increased mineralization of hMSCs led to bone growth.
Figure 12.5 Coating and doping of the beta-TCP scaffolds.
312 Chapter 12
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Samarasinghe et al., 2014 formulated nano-formulation C-CP-NCs to de-
liver CQ extracts. They used mice as the animal model and demonstrated the
effect of the extract to ameliorate OA disease progression by inducing
chondral protective and disease-modifying activity. A signaling mechanism
modulated by alginate enclosed C-CP-NCs in chondrocyte was proposed.
This study also established the non-toxic and biodegradable nature of the
formulation with superior therapeutic activity.
102
A phytochemical scaffold was fabricated by mixing CQ extract with a
chitosan/Na-carboxymethyl cellulose blend via a lyophilization technique for
bone regeneration. It was found that the addition of CQ increased the
compression modulus of the composites and were suitable for bone re-
generation. The fabricated scaffolds exhibited an osteoinductive property
suggesting their potential application in bone tissue engineering.
103
Thongtham et al., in 2020 fabricated the bone tissue engineering compos-
ite scaffolds using chitosan/collagen/hydroxyapatite biopolymers to en-
capsulate CQ loaded polycaprolactone nanoparticles prepared by the double
emulsion method. The prepared biopolymer scaffold was shown to prevent
the sudden release of CQ and showed an improved release profile along with
a prolonged release rate due to a double sustained mechanism. The fabri-
cated scaffolds were also found to help in osteoblast proliferation and were
non-toxic to MC3T3-E1 murine osteoblasts.
104
12.6 CQ Safety Assessment in Animal Models and in
Clinical Studies
CQ stems, extracts, and powders are commonly used in traditional systems
of medicine and also consumed by the local community as a general health
tonic, to promote bone and tissue healing, to treat infections, and to pro-
mote weight loss and management. Scientific reports established its toxicity
level by using animal models and they support the use of CQ in the health
system. Some studies concluded that CQ appears to be very safe at the doses
commonly used in toxicity studies. Some other studies recommended fur-
ther standardization and clinical toxicity data.
105
The sub-chronic toxicity of
CQ stem powder was evaluated in Wistar rats by Attawish et al., in 2002. They
found that oral treatment with a CQ powder suspension in water at four
different concentrations (0.03, 0.3, 3.0, and 3.0 g per kg body weight per day)
for three months did not reveal any significant changes in hematological
and biochemical parameters, body weight before and after the study, as well
as in histopathological studies indicating it to be possibly safe at a 1000-fold
therapeutic dose in humans.
106
Kothari et al., in 2011 investigated the sub-chronic toxicity of CQR-300 in
Sprague Dawley rats for 90 days. CQR extract at doses (0–2500 mg per kg body
weight) did not produce any clinical toxicity, including mortality, change in
body weight, microscopic or macroscopic abnormalities, and changes in
hematological, biochemical, urine analysis, and ophthalmological parameters.
Phytochemical and Pharmacological Aspects of Cissus quadrangularis in Ayurveda 313
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Furthermore, results of an Ames assay of mutagenicity indicated that CQR-300
is not genotoxic. Based on the results of this study, it was suggested that CQ
extract up to a dose of 150 g per person per day won’t produce any adverse
effect and hence is very safe for human consumption.
107
Similarly, adminis-
tration of a standardized phytoestrogen-rich fraction of CQ to the rats in a eight
week-long study also showed no signs of toxicity.
20
In a sub-acute toxicity study
of a CQ hexane extract, chronic administration of it did not produce any
physiological and pathological changes as compared to normal rats.
73
In an-
other study, the sub-chronic toxicity of enteric film coated tablets containing
CQ extract was evaluated in Wistar rats. Animals were given tablets containing
7.14–714 mg per kg of CQ extract for 90 days. None of the animals showed any
sign of dose related toxicity on hematological parameters, serum chemistry,
urine analysis, and macroscopic abnormalities.
83
Kavitha et al., in 2018 dem-
onstrated that an aqueous extract of the whole plant of CQ at a dose of 3000 mg
per kg bodyweight upon administration to Swiss albino mice for 14 days did
not produce any toxicity.
108
Contrary to preclinical trials, a clinical case report
stated that herbal capsules containing CQ extract induced thrombocytopenia
in a renal allograft patient. This patient was under an immunosuppressive
regimen for his ailment and suddenly reported severe thrombocytopenia after
consuming CQ herbal capsules. A prompt improvement in the condition of the
patient has been observed after the withdrawal of the CQ herbal capsules.
Hence the physician related this effect to the CQ extract for thrombocytopenia
and advised avoiding CQ herbal capsules for patients under immuno-
suppressive therapy as there is no supporting evidence about the drug inter-
actions and also a lack of available data about the adverse drug reaction (ADR).
The data from the acute and sub-chronic animal studies
20,106,107
indicate
that daily use of CQ stems powder or extract for 90 days is not associated
with any unwanted side effects confirming its safety. However, there is a
scarcity of data on their long-term clinical use in humans.
12.7 Intellectual Property Rights Related to CQ
Granted patents and applications related to the use of CQ for various dis-
orders are presented in Table 12.3.
12.8 Conclusion and Future Perspectives on CQ
Clinical Use
Existing systematic and meta-analyses of reviews related to CQ suggest that
CQ is possibly safe and more effective in the management of bone fractures,
bone related disorders, and obesity/weight loss management, but multiple
clinical evidence is lacking. The effect of CQ in obesity/weight loss
management is inconclusive because of the lack of multiple evidence. Meta-
analysis reviews suggest that CQ is effective in combination with other
ingredients for the management of obesity, but the effectiveness is in-
conclusive. Many studies reported the effect of CQ in bone fractures, but the
314 Chapter 12
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Table 12.3 Summary of patents/patent applications related to CQ.
S. no. Patent details Inventors Title of invention Description
1. US20050048141A1
2007-01-19
Brian Cortes
Lal Hingorani
Vijay Thawani
Use of the plant CQ or its extracts
for osteo-health including
prevention or mitigation of
degenerative disease, fracture
healing, and anabolic increase
in osteo tissues
This invention describes the composition
and preparation of a nutritional product or
a dietary supplement containing CQ or its
extracts to restore bone health and in the
treatment of osteo-health related
conditions.
2. WO2008081233A3
2009-11-19
Villoo M Patell CQ plant extracts for treating
osteoporosis and the extraction
process thereof
It is related to the preparation of CQ extracts
and its efficacy in the treatment of
osteoporosis.
3. US8318213B2
2012-11-27
Julius E. Oben Plant extract mixtures and their
uses
A composition comprising an effective amount
of CQ leaves and stems extract claims to be
beneficial in weight loss in obesity.
4. WO2020089944A1
2020-05-07
Nilima Mohanty;
Somashekara
Nirvanashetty;
Sanjib K. Panda
Vivek A Parachur
Process for preparing a herbal
extract and compositions
thereof
It describes a novel process for the isolation
of selective compounds and resins in
specific concentrations for preparation of a
herbal extract and compositions comprising
the same.
5. US2008166419A1
10-07-2008
Scott F. Sones Krill oil compositions This invention claims a composition
containing CQ standardized to 5%
ketosteroids which is beneficial in reducing
one or more risk factors associated with
cardiovascular diseases.
6. WO2020236023A1
2020-11-26
Anton L.
Prozumentov
Georgiy V. Sarychev
Vera K. Vedmina
Ivan S. Martynenko
Dietary supplement formulation
for weight loss
It is related to composition of a dietary
supplement containing various plant
extracts including CQ stalks for weight loss.
7. EP2941261B1
24-07-2019
Ganga R. Gokaraju
Rama R. Gokaraju
Venkata K. R. R.
Gokaraju
Synergistic curcuma and cissus
compositions for enhancing
physical performance and
energy levels
It claims a synergistic dietary supplement
compositions having extracts (20% to 80%)
of CQ and Curcuma longa for improving
physical performance and mental alertness.
Phytochemical and Pharmacological Aspects of Cissus quadrangularis in Ayurveda 315
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Table 12.3 (Continued)
S. no. Patent details Inventors Title of invention Description
Kiran Bhupathiraju
Trimurtulu Golakoti
Krishanu Sengupta
Venkata K. R. Alluri
8. US7582316B1
01-09-2009
Jacob Geissler Ketone-steroid extracts from the
CQ plant and methods thereof
This invention discloses composition and
preparation of ketone steroid extract of the
roots of CQ which can be used to promote
lean body mass, treat damaged or diseased
connective tissue, and provide an analgesic
effect while reducing cortisol.
9. US10542989B2
28-01-2020
Ganga R. Gokaraju
Rama R. Gokaraju
Venkata K. R. R.
Gokaraju
Kiran Bhupathiraju
Trimurtulu Golakoti
Krishanu Sengupta
Venkata K. R. Alluri
Synergistic dietary supplement
compositions for enhancing
physical performance and
energy levels
It discloses the composition of a novel
synergistic dietary supplement containing
at least two plant extracts to enhance the
body energy levels that can augment
physical and mental health.
10. US7935367B2
03-05-2011
Marvin Heuer
Ken Clement
Shan Chaudhuri
Compositions and methods for
increasing adipose metabolism,
lipolysis, or lipolytic
metabolism via thermogenesis
This invention discloses a composition of a
dietary supplement containing extracts of
CQ (0.05 g to about 0.3 g) for enhancing
lipid metabolism.
11. WO2007101066A3
2008-10-30
Shil C Kothari Methods and compositions for
improving cardiovascular risk
factors and metabolic risk
factors that cause syndrome x
This invention discloses the beneficial use of
300 mg CQ extract per day for six weeks in
obese patients such as weight loss, appetite
suppression, decrease in LDL, and blood
sugar and triglyceride levels.
316 Chapter 12
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12. WO02085394A1
2002-10-31
Bakulesh M.
Khamar, Vandana
D. Mody
Kalpesh K. Pandya
The process of preparing the
topical anti-inflammatory/
analgesic preparation
This invention provides a method of
preparation of a synergistic herbal
preparation containing CQ and Costus
species as main ingredients along with
other volatile oils. The preparation is
claimed to be effective in relieving painful
and inflammatory conditions.
13. WO2008028268A1
2008-03-13
Marvin Heuer
Ken Clement
Shan Chaudhuri
Diet supplement for causing
weight loss comprising CQ,
green tea, and caffeine
It claims a dietary supplement containing CQ
stem and leaf extract (0.01 g to 1 g), green
tea leaf extract, and anhydrous caffeine
(max 0.3 g) for causing weight loss.
14. WO2010054469A9
2010-07-08
Marvin A. Heuer
Ken Clement Megan
Thomas
Jason Peters
Appetite suppressing weight
management composition
comprising Caralluma
fimbriata, Hoodia gordonii, and
CQ extracts
It claims a method of preparation of a
composition containing five ingredients to
reduce body weight and suppress appetite.
15. US7659313B2
2010-02-09
Julius E. Oben Methods and related
compositions using specific
flavonoids and indanes to
reduce weight and inhibit
lipase, alpha-amylase, and
alpha-glucosidase activity in
mammals
It claims a composition (50–200 mg per day)
prepared by using flavonoids and indanes
extracted from the stem and leaves of CQ for
weight loss. Selected flavonoids or indanes
include 3-O-rhamnopyranosyl kaempferol;
3-(4-hydroxybenzylidene)-2-(2,5-
dihydroxyphenyl)-1-(4-
hydroxyphenyl)indane-4,6-diol; quercitrin;
rhamnitrin; rhamnocitrin; quercitrin-3-O
00
-
acetate; parthenocissin A, etc.
Phytochemical and Pharmacological Aspects of Cissus quadrangularis in Ayurveda 317
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depth of the study and the plausible mechanism is still not clear. Several
clinical studies reported the pharmacological activity of CQ but they have
their own limitations like small sample size, poor design and methodo-
logical incompliances, inadequate reporting, and medical interventions.
Sawangjit et al., in 2017 pooled the effects of CQ in different dosage forms,
dose and clinical conditions especially for bone related ailments. They
concluded that future studies are warranted to determine whether the dose,
dosage form, and the clinical condition correlates to the pharmacological
effects of CQ and also the combinational therapies for better therapeutic
management.
109
They also mentioned that most of the included studies in
meta-analyses (six of nine) indicated that CQ is safe for human consumption
for a period of 1 to 10 weeks. However, there is limited data on the long-term
safety and clinical use in different ailments and future research reports with
strong evidence are recommended.
109
Extra randomized clinical trials are
needed to establi sh the safety and efficacy of CQ for the management and
treatment of various ailments.
This chapter consolidated the significance of CQ phytochemical con-
stituents and pharmacological actions with the hope that comprehensive
information about CQ is essential for future scientific investigations in order
to generate novel therapeutically effective lead molecules. The next level of
phytochemical investigations should be carried out to isolate potent mol-
ecules from different fractions of the CQ extract and expedite the develop-
ment of medicine. In vitro and in vivo preclinical studies are extensively
reported, but still a large gap exists between the relationship of active
phytoconstituents – targets – molecular mechanisms – therapeutic effects.
Substantial clinical study to support the pharmacological activity and
human safety of CQ is imperative and inevitable. With the advancement of
modern analytical and molecular techniques coupled with the conventional
methods, future research should make a significant contribution towards
the development of newer therapeutic agents from CQ. Several daunting
challenges need to be addressed effectively for the development of lead
molecules from medicinal plants through collaborative efforts of anthro-
pologists, ethnobotanists, phytochemists, pharmacists, and physicians.
Future studies should focus on bridging the gap between in vitro/in vivo
and clinical effects to assure the therapeutic efficacy of this incredible
herbal plant.
Abbreviations
AgNPs Silver nanoparticles
ALKP Alkaline phosphatase
AuNPs Gold nanoparticles
BMI Body mass index
cAMP Cyclic adenosine monophosphate
CAT Catalase
CFA Complete Freunds Adjuvant
318 Chapter 12
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COX Cyclooxygenase
Cp Cysteine protease
CQ Cissus quadran gularis
CuONPs Copper oxide nanoparticles
DEXA Dual-energy X-ray absorptiometry
DNA Deoxy ribose nucleic acid
DPPH 2,2-Diphenyl picrylhydrazyl
ER Estrogen receptor
GABA Gamma amino butyric acid
GAE Gallic acid equivalent
GPx Glutathione peroxidase
GSH Reduced glutathione
GST Glutathione S-transferase
H
2
O
2
Hydrogen peroxide
HDL High density lipoprotein
hMSCs Human mesenchymal stem cells
HO-1 Heme oxygenase-1
hWA Human white adipocytes
IGF Insulin-like growth factor
IL Interleukin
iNOS Inducible nitric oxide synthase
IR Infra-red
LDL Low densi ty lipoprotein
LOX Lipooxygenase
LPS Lipopolysaccharide
MD-OADs Muscular dystrophy during osteoarthritic
disorders
MTT (3-(4,5-Dimethylthiazolyl-2)-2,5-diphenyl-
tetrazolium bromide)
NMR Nuclear magnetic resonance
NO Nitric oxide
Nrf Nuclear transcription factor
NSAID Non-steroidal anti-inflamm atory drugs
OA Osteoarthritis
OPG Osteoprotegrin
PGE2 Prostaglandin E2
PLLA Poly-
L-lactic acid
RANKL Receptor Activator of Nuclear Factor
K Ligand
ROS React ive oxygen species
SOD Superoxide dismutase
StAR Steroidogenic acute regulatory protein
TCP b-Tricalcium phosphate
TEM Transmission electron microscopy
TNF-a Tumor necrosis factor-a
TRAP Tartrate resistant acid phosphatase
Phytochemical and Pharmacological Aspects of Cissus quadrangularis in Ayurveda 319
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